SynopsisInformation on spatial correlation in the tangent direction along electron microscope images of filamentous molecule is shown to be obtainable by the analysis of statistical fluctuations in curvature, yielding an absolute measure of the persistence parameter a,i ,,,.The relationship of amicro, a local, microscopic parameter, to the persistence length introduced by Kratky and Porod is discussed. The hypotheses underlying the assumed theoretical model concern (1) the shape of the angle distribution, assumed to be Gaussian; (2) the passage from a three-to a two-dimensional situation, which is supposed to occur by deformation of the flexible chain in a manner that preserves the memory of the spatial correlation in orientation (except for the blocking of one degree of freedom); and (3) the adsorption conditions, which should meet the equilibrium requirement as closely as possible. The analytical method has been checked on computer simulated "Gaussian" molecules: the study of the simulated sample was essential in solving the problems connected with minimum statistics requirements and the effect of the reading error. Experimental images obtained for T2 DNA fragments a t different ionic strengths by Kleinschmidt's adsorption technique have been analyzed by means of an automatic flying spot digitizer, the "Precision Encoder and Pattern Recognition." The results show that adsorbed molecules do in fact "remember" the rigidity they possessed in solution and that the Gaussian hypothesis is well verified. Consequently, the slopes of log a ( 1 ) or @ ( l ) may be used indifferently in the estimate of amicro The dependence of this parameter on ionic strength in the range explored shows the expected behavior.

A kinetic study of the alkaline transition of DNA, in clearly defined physico-chemical conditions, is presented, which allows us to identify, within the alkaline transition region, different pH ranges, corresponding to different ratelimiting factors. This analysis brings into consideration three distinct intervals of time which characterize the whole process, namely the time necessary for full hyperchromicity to be reached, the time required for strand separation in the case of a single DNA molecule, and the time for complete denaturation to be reached in the case of a DNA solution.THE RESULTS OBTAINED FROM ULTRACENTRIFUGAL, AND SPECTROPHOTOMETRIC MEASUREMENTS, INVOLVING RAPID MIXING EXPERIMENTS, SEEM TO INDICATE THE FOLLOWING CONCLUSIONS: whereas, in the lower pH ranges considered within the transition region, the denaturation process is limited by the first time constant, this same constant becomes extremely short at higher pH. On the other hand the fact that, in the higher pH range, the second and third time constants do not coincide (the time to unwind a single T2 DNA molecule being at least one order of magnitude shorter than the time required for bulk denaturation to be reached) suggests that in this pH range the overall denaturation rate is limited by a statistical process governing the initiation of unwinding.These observations are discussed in terms of a model in which the unwinding energy is given by the electrostatic repulsions which originate in the deprotonated DNA molecule. The model itself suggests some experiment which seem to confirm it.

A fragment of Plasmodium berghei DNA was cloned using a technique designed to select for telomeric sequences. The cloned fragment recognizes Bal31‐sensitive bands in P. berghei genomic digests. It contains at its distal end at least 70 tandem repeats of the heptanucleotide sequence CCCTGAAA. The presence of natural single strand discontinuities in the telomeric regions of P. berghei DNA is demonstrated by the selective incorporation of deoxyribonucleoside triphosphates in the absence of DNase. The number of copies of the cloned sequence present in each genome agrees with an estimate of 6‐12 chromosomes per nucleus.

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